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J Autoimmun ; 34(1): 59-65, 2010 Feb.
Article in English | MEDLINE | ID: mdl-19683415

ABSTRACT

A diagnostic subgroup of AIH type 1 is characterized by specific serum antibodies against soluble liver protein. The respective autoantigen was named SLA/LP/tRNP((Ser)Sec), after three homologous recombinant polypeptides were isolated from expression gene libraries. We analyzed human cultured liver cells for the human homologue of recombinant SLA/LP/tRNP((Ser)Sec) by antigen purification. In addition, a monoclonal antibody was generated against recombinant SLA-p35, a truncated recombinant SLA-reactive polypeptide. With a positive patient serum, immune affinity chromatography was performed on the 52 kD-SLA main antigenic determinant pre-enriched by ion exchange chromatography. By mass spectrometry, the 52 kD-SLA/LP/tRNP ((Ser)Sec) autoantigen was unambiguously identified in the purification product. The identity of the recombinant SLA-p35 and its human homologue was further confirmed by a specific signal of the anti SLA-p35 monoclonal antibody with purified human SLA/LP/tRNP((Ser)Sec). The 48 kD-SLA species frequently comigrating in SLA-immunoblotting however was not identified by either approach. We conclude that the native counterpart of recombinant tRNP((Ser)(Sec)) indeed is detectable with a molecular weight of 52 kD in soluble liver extract of human cells as the major antigenic component of SLA/LP/tRNP((Ser)Sec).


Subject(s)
Antibodies, Monoclonal/metabolism , Autoantigens/metabolism , Hepatitis, Autoimmune/metabolism , Hepatocytes/metabolism , RNA, Transfer, Amino Acyl/metabolism , Amino Acid Sequence , Autoantigens/blood , Autoantigens/genetics , Autoantigens/immunology , Cell Extracts , Cell Line, Tumor , Chromatography, Affinity , Cloning, Molecular , Hepatitis, Autoimmune/blood , Hepatitis, Autoimmune/diagnosis , Hepatitis, Autoimmune/immunology , Hepatocytes/immunology , Hepatocytes/pathology , Humans , Mass Spectrometry , Molecular Sequence Data , Peptide Fragments/blood , Peptide Fragments/genetics , Peptide Fragments/immunology , Peptide Fragments/metabolism , RNA, Transfer, Amino Acyl/blood , RNA, Transfer, Amino Acyl/genetics , RNA, Transfer, Amino Acyl/immunology , Recombinant Proteins/blood , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/metabolism
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